Hi,
I am planning on seeding rat MSCs onto 6 well plates with coverslips attached on the bottom. I then want to use stains such as Alizarin Red and Von Kossa to see the level of osteogenic differentiation. Is there a way of quantifying the stains after placing the coverslips onto microscope slides to view the stain or will quantification have to happen with the coverslip in the plate?
The quantification of staining methods is a bit complicated. You can e.g. put your slides on a normal scanner. The von Kossa staining is based on the exchange of Calcium ions with silver ion the last appear black after treatment with Pyrrogallol. Scanning the stained slides allows a quatification of the black area.
For direct quantification of Calcium and Phosphat I would recommend the Calcium CPC Kit (Greiner) and the Phosphorus UV Liquirapid Test (Greiner). This test allows after HCL treatment the quatification of Calcium and Phosphat via photometry.
For quatification Alkaline Phosphatase you could an activity test.
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